有效及快速的在不同來源(天然或合成) 的複雜基質中檢測出生物活性物質的研 究，在藥物開發上相當具有潛力。在這些生物活性物質中，能與核酸產生交互作用的 物質為研究的重要標的之一。近年來表面電漿共振技術於生物分子交互作用分析的發 展，可以在不須標記的情況下，即時的檢視分子間的交互作用。本計畫的目的是建立 以SPR 技術為基礎的感測平台，應用於篩選含有能與DNA 結合之物質的植物、中草 藥。本計畫所用的DNA 探針為模仿人類免疫不全病毒(Human immunodeficiency type 1 virus, HIV1)長端點重覆序列(long terminal repeat, LTR)的Sp1，NF-kB 及TFIID 結合位 置的序列。此外，以石蒜科的孤挺花及毛茛科的黃連作為模式系統，因為這兩種植物 都含有大量生物鹼，而生物鹼已知能與DNA 產生結合。首先以層析法區分植株的粗抽 出物，各區分再以本計畫提出之感測平台進行能與DNA 分子結合的生物活性物質的篩 選。而後以生物分析及光譜分析法平行驗證此感測平台的可行性。最後將此技術應用 於可食植物的篩選，以期能有效開發植物資源，作為發展具抗癌、抗病毒功能的藥草 的新標的。
The efficient and rapid detection of bioactive compounds in complex matrices of different origins (natural or synthetic) is a key step in the discovery of molecules with potential application in therapy. Among them, molecules able to interact with nucleic acids can represent important targets. The recent development of biosensor technologies for biomolecular interaction analysis enables monitoring of a variety of molecular reactions in real-time by surface plasmon resonance (SPR). In this study, we aim to demonstrate the proposed DNA biosensor, based on SPR transduction, is potential as a new platform for bioactive compounds screening. The target DNA sequence are synthetic oligonucleotides mimicking the Sp1, NF-kB, and TFIID binding sites of the long terminal repeat of the human immunodeficiency type 1 virus. Fractions obtained by chromatographic separation of the extract of Hippeastrum hybridum (Barbados Lily) and Coptis chinensis Franch, plants containing alkaloids having intercalating properties, are examined for their ability to interact with the immobilized DNA probe. The presence of alkaloids in the investigated fractions obtained from the extract is evaluated, in parallel, by conventional bioassay and spectrophotmetric analysis. Furthermore, edible plants are screened for their DNA-binding activity based on the proposed platform. This work demonstrates the potential of applying the biosensor approach in the search for bioactive compounds from plants.
